聚天冬酰胺键合胰蛋白酶亲合手性固定相的初步研究—Ⅰ

Studies on the Poly-Aspartic Trypsin Chiral Stationary Phase——Ⅰ

目的 :将胰蛋白酶通过N ,N′-羰基二咪唑以共价结合的方法键合于双亲性载体聚 - (羟丙基 ,丙基 ) -DL -天冬酰胺材料上 ,制成亲合手性固定相。方法 :用X -衍射、扫描电镜等方法进行了表征。对固定相进行了pH、温度、时间及热稳定性等优化条件的选择。结果 :固定相在 37℃～ 45℃能稳定存在 ,在pH 7 4～ 10 6 ,37℃条件下有稳定的酶活性。结论 :该固定相有可能成为蛋白分离的固定相。

Objective:A new chiral stationary phase,trypsin-poly-(hydroxypropyl,propyl)-aspartamide,(Try-PHPPA),was synthesized.Methods:The active protein,trypsin,was conjugated to the carrier by the cross-linking,1,1′-carbodi-imidazole at room temperature.The chiral stationary phase was characterized by X-ray and SEM,and the activity and characteristics of Try-PHPPA were measured.Results:It showed that the protein,coupled with the carrier has higher thermal stability than the free protein,the stability temperature is range from 37 ℃ to 45 ℃.It has stable protein activity at weak buffer solution,pH 74～106,37 ℃.The Try-PHPPA also kept its protein activity for 72 h under the experiment conditions.Conclusion:It may be a useful chiral stationary, and further experiments have been carrying out.